LOCUS Exported 5429 bp ds-DNA circular SYN 07-JAN-2018
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..5429
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..285
/gene="araBAD"
/label=araBAD promoter
/note="promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction (Guzman et al., 1995)"
RBS 312..334
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 354..371
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 375..407
/codon_start=1
/product="leader peptide from bacteriophage T7 gene 10"
/label=T7 tag (gene 10 leader)
/note="promotes efficient translation in E. coli"
/translation="MASMTGGQQMG"
CDS 420..440
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
CDS 447..1166
/codon_start=1
/product="enhanced monomeric variant of CFP (Goedhart et
al., 2012)"
/label=mTurquoise2
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 447..1166
/codon_start=1
/product="enhanced monomeric variant of CFP (Goedhart et
al., 2010)"
/label=mTurquoise
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
terminator 1382..1468
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 1560..1587
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 1606..1697
/gene="bla"
/label=AmpR promoter
CDS 1698..2558
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 2600..3055
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
rep_origin 3166..3754
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 3940..4080
/label=bom
/note="basis of mobility region from pBR322"
CDS complement(4525..5403)
/codon_start=1
/gene="araC"
/product="L-arabinose regulatory protein"
/label=araC
/translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
EKVNDVAVKLS"
ORIGIN
1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
241 atactacctg acgcttttta tcgcaactct ctactgtttc tccatacccg tttttttggg
301 ctagaaataa ttttgtttaa ctttaagaag gagatataca tatgcggggt tctcatcatc
361 atcatcatca tggtatggct agcatgactg gtggacagca aatgggtcgg gatctgtacg
421 agaacctgta cttccagggc tcgagcatgg tgagcaaggg cgaggagctg ttcaccgggg
481 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
541 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
601 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gtcctggggc gtgcagtgct
661 tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
721 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
781 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
841 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacttcagc gacaacgtct
901 atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaaca
961 tcgaggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
1021 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccaagctg agcaaagacc
1081 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
1141 tcggcatgga cgagctgtac aagtaagaat tcgaagcttg gctgttttgg cggatgagag
1201 aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat
1261 ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa
1321 cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca
1381 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
1441 ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca
1501 acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca
1561 gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttttgt ttatttttct
1621 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
1681 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
1741 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
1801 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
1861 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
1921 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
1981 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
2041 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
2101 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
2161 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
2221 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
2281 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
2341 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
2401 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
2461 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
2521 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
2581 atatacttta gattgattta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
2641 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
2701 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
2761 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
2821 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
2881 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
2941 ggctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
3001 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aatttaaaag
3061 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
3121 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
3181 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
3241 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
3301 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
3361 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
3421 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
3481 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
3541 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
3601 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
3661 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
3721 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
3781 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
3841 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
3901 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
3961 tacgcatctg tgcggtattt cacaccgcat ctggtgcact ctcagtacaa tctgctctga
4021 tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg
4081 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
4141 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
4201 tcaccgaaac gcgcgaggca gaaggagatg gcgcccaaca gtcccccggc cacggggcct
4261 gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc
4321 ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg
4381 gccacgatgc gtccggcgta gaggatctgc tcatgtttga cagcttatca tcgatgcata
4441 atgtgcctgt caaatggacg aagcagggat tctgcaaacc ctatgctact ccgtcaagcc
4501 gtcaattgtc tgattcgtta ccaattatga caacttgacg gctacatcat tcactttttc
4561 ttcacaaccg gcacggaact cgctcgggct ggccccggtg cattttttaa atacccgcga
4621 gaaatagagt tgatcgtcaa aaccaacatt gcgaccgacg gtggcgatag gcatccgggt
4681 ggtgctcaaa agcagcttcg cctggctgat acgttggtcc tcgcgccagc ttaagacgct
4741 aatccctaac tgctggcgga aaagatgtga cagacgcgac ggcgacaagc aaacatgctg
4801 tgcgacgctg gcgatatcaa aattgctgtc tgccaggtga tcgctgatgt actgacaagc
4861 ctcgcgtacc cgattatcca tcggtggatg gagcgactcg ttaatcgctt ccatgcgccg
4921 cagtaacaat tgctcaagca gatttatcgc cagcagctcc gaatagcgcc cttccccttg
4981 cccggcgtta atgatttgcc caaacaggtc gctgaaatgc ggctggtgcg cttcatccgg
5041 gcgaaagaac cccgtattgg caaatattga cggccagtta agccattcat gccagtaggc
5101 gcgcggacga aagtaaaccc actggtgata ccattcgcga gcctccggat gacgaccgta
5161 gtgatgaatc tctcctggcg ggaacagcaa aatatcaccc ggtcggcaaa caaattctcg
5221 tccctgattt ttcaccaccc cctgaccgcg aatggtgaga ttgagaatat aacctttcat
5281 tcccagcggt cggtcgataa aaaaatcgag ataaccgttg gcctcaatcg gcgttaaacc
5341 cgccaccaga tgggcattaa acgagtatcc cggcagcagg ggatcatttt gcgcttcagc
5401 catacttttc atactcccgc cattcagag
//